CD8+ T cells transduced with PSCA-28t28Z (second generation CAR) and isolated 30 days after adoptive transfer into mice bearing HPAC-derived pancreatic tumor. Human peripheral blood mononuclear cells were stimulated with anti-CD3 antibody (OKT3) in presence of IL-2. Two days post-stimulation, T cells were transduced with retroviral vectors encoding PSCA-28t28Z. Cells were cultured for 2 weeks in presence of IL-2 and then transfered into 4-5-week-old male NSG mice. Subcutaneous xenografts were generated by injection of HPAC cells. Once tumors became palpable, mice were treated with CD8+ T cells expressing PSCA-28t28Z. Untransduced CD4+ cells from the same donor were given to each mouse for cytokine support. Spleen-resident human CD8+ T cells were isolated 30 days later using the CD8 MicroBeads (post-infusion samples).

T-cell isolation study 11 / T-cell isolation study 6

CD4+ resting memory T-cell were isolated from peripheral blood buffy coat of healthy donors after storage for 24 hours at 20°C. The cell fraction was first enriched via density gradient centrifugation with LSM 1077 Ficoll and than FACS sorted as CD3+/CD4+/CD45RO+/CD45RA- population.

Control

T-cell isolation study 6

CD4+ resting memory T-cell were isolated from whole peripheral blood of healthy donors with no delay. The cell fraction was first enriched via density gradient centrifugation with LSM 1077 Ficoll and than FACS sorted as CD3+/CD4+/CD45RO+/CD45RA- population.

Oligodendrocyte progenitor cells (OPC) isolated from cortical tissue, which was obtained from patients with epilepsy, but without any manifested brain cancer. OPC were isolated using magnetic-activated cell sorting (MACS) with antibodies against A2B5 antigen.

Oligodendrocyte progenitor cells (OPC) isolated from cortical tissue, which was obtained from patients with epilepsy, but without any manifested brain cancer. OPC were isolated using magnetic-activated cell sorting (MACS) with antibodies against A2B5 antigen.